WO2017010858A1

WO2017010858A1 - Composition based on extracts of heliopsis longipes and propolis for the control and/or elimination of ticks

Info

Publication number: WO2017010858A1
Application number: PCT/MX2015/000104
Authority: WO
Inventors: Beatriz FLORES SAMANIEGO
Original assignee: Flores Samaniego Beatriz
Priority date: 2015-07-15
Filing date: 2015-07-15
Publication date: 2017-01-19
Also published as: CO2017007327A2

Abstract

The invention relates to an insecticide composition based on natural extracts for the control and/or elimination of ticks in livestock and/or in domestic animals, characterised by comprising: a) 50 to 65% Heliopsis logipes extract; b) 5 to 20% propolis extract; and c) 30% dermal application vehicle, all of which must add up to 100%. The mixture of the composition has a synergetic effect guaranteeing the control of adults, nymphs, pupae, larvae and eggs, in particular in adult females, by altering their capacity for ovipositioning and also affecting the possibility of the eggs hatching and the survival of the larvae, in particular for all the ticks of veterinary importance of the Ixodidae family and the Argasidae family.

Description

Composition based on extracts from H @ íiopsis lorsgipes and Propoüs for @S contro! and / or eradication of ticks

Field of the Invention

The present invention relates to a composition based on natural extracts of Heüopsis íongipes and Propolis for the control and / or elimination of ticks, where e! Heüopsis Iongipes extract has effects of paralysis and toxicity on the ticks, acting in synergy with the Propolis extract which has active principles of high toxicity against insects, for use in cattle and / or domestic animals.

Before the invention

The tick is one of the most economically important ectoparasites worldwide, due to the losses it causes in the production of cattle, goats, sheep and horses. Direct consequences of tick parasitization are weight loss due to reduced food intake and due to toxins and skin irritation, anemias caused by blood loss, transmission of hemoparasites (Babesia bígemina, B. bovis and Anaplasnja margínale ) and the considerable depreciation of the skins due to the perforations produced by the pickets. In addition, these perforations allow infection by bacteria, development of dermal mycoses and entry of fly larvae (myiasis). By injuring the skin to suck blood, many species of ticks can transmit various pathogens such as viruses, bacteria, riches and protozoa. This can lead to acute and / or chronic diseases such as piroplasmosis, Lyme disease, ehrlichiosis, tberyleriosis, Chagas disease, Q fever and uraemia, and even death. Of the estimated 1,000 million head of cattle in the world, between 70 and 80% live in tropical and subtropical countries, in Sos that the tick is active throughout the year.

Tick infestations can be very intense, so it is not uncommon to find animals parasitized by hundreds and even thousands of specimens, which logically It multiplies the damages, both direct and derived from the transmission of diseases whose prophylaxis and treatment is an important problem facing contemporary veterinary medicine.

In animals, damage can be very showy and flashy. Sos are more frequent as follows: a) tissue destruction caused by Sos mouth appendages of ticks and the inflammatory reaction that occurs in response to the bite; b) blood exposure, which can cause acute anemias in animals with intense infestations; c) paralysis caused by salivary toxins of some species of ticks, such as the neurotoxin called hofocyclotoxin of the Australian tick Ixodes oiocyclus, a single female of this species can cause paralysis and death of an animal; and d) toxicosis, such as s eating sickness or sweat ios disease, caused by salivary toxins of the African species Hyalo a truncatum in ruminants, this disease presents with eczemaphic skin lesions and hypersecretion of exudates and causes mortalities greater than 75% in animals young boys.

From the point of view of damage and power! Transmitting diseases, on a global scale, nine genera of ticks Ixodidae {Ambiyo ma spp., Boophüus spp., Dermacentor spp., Haemaphysalis spp., Hyalomma spp., ixodes and R ipicephalus) and Argasidae (Omithodorus spp. and Otogasodpius Otop. spp.)

In recent years the problem of ticks has increased and the poor management of its control has created serious problems of multiple resistance to the different chemicals with which they are fought.

The most used strategy for tick control consists in the application at specific intervals of ixodicides on the body of infested animals. Most tick products contain active contact ingredients (organophosphates, pyrethroids, amidines and fipronsl) and are available as concentrates for immersion or spray baths or as "pour-on" ready for use. They are effective against larvae, nymphs and adults. Many of them also control other parasites of cattle such as flies, lice, mites, etc. However, its irrational use has caused the generation of tick strains resistant to the action of these chemicals and even combinations of them.

Resistance is a phenomenon that occurs in any type of population, whether insects, arthropods, bacteria, etc., when it is subjected to the constant and prolonged action of some substance, so that some individuals are generating resistance to Certain doses of the products. Once this capacity has been developed, individuals survive the treatments and when they reproduce, their offspring will also be resistant. In addition, both the discharge or elimination of Sos garrapaicic baths and Sa of the containers containing these products pollute the environment. In response to this problem and seeking to reduce the costs involved in the creation of new biologically active molecules, the insecticidal properties of plañías have been investigated, which by popular order have been recognized in ethnobotanical studies as such.

From the need to find an efficient, natural tick that allows to reduce the use of harmful chemicals, a new alternative arises to make a mixture of natural extracts, such as cniícuague extract (Heüopsís Songipes) and propoleo (Propoiis ), both with high biological activity, which allow the control and / or elimination of ticks in cattle, sheep, goats and domestic animals.

Document CN10359922Q describes a composition for human acariasis containing propolis mixed with four roots of Asian origin, carbonized cabelium, four plants (three of Asian origin and one of Iberian origin) and two fruits of Asian origin; whose composition although it contains propolis, differs totally from the one presented here.

Document CN102670939 describes a composition for acariasis in cattle containing propolis mixed with three roots of Asian origin, three plants of Asian origin, two plants of Euro / Asian origin, a world-class plant, dried orange peel and calcium sulfate; whose composition although it contains propolis, differs totally from the one presented here, Document CN 102670863 describes a composition for pet acariasis containing propolis mixed with sulfur, hunger, a miner! of arsenic, two plants of world origin, a root of Asian origin and an organic compound of the group of érpenos; whose composition although it contains propolis, differs totally from the one presented here.

Ia is erseiórs description

The present invention relates to a composition based on extracts of Heiiopsis longipes and Propolis, for the control and / or elimination of ticks, in which the extract of Heiiopsis longipes has effects of paralysis and toxicity on ticks, acting in synergy with the Propoüs extract and which has active ingredients of high toxicity against insects; For use in livestock and pets.

The composition of the present invention for the control and / or elimination of ticks, comprises: a) A natural extract of Heiiopsis longipes from 50 to 85% v / v

b) A natural extract of Propoüs from 5 a! 20% v / v and

c) A 30% dermal vehicle v / v

All this must add up to 100%

The extract of chilcuague (Heiiopsis longipes) in the composition works through its main active ingredient, afinin, which is the aicamide responsible for the main biological effects observed. It has insecticidal activity causing paralysis and toxicity against insects,

Propolis extract (Propolis) in the composition works through curnarins that are toxic to ticks, synergizing with chilcuague extract. Additionally, the propolis extract in the composition provides antibiotic protection to the animals through the action of its amphibious and antifungal components. The dermal vehicle is selected from giicerina, mineral oil, petroleum jelly, etc. and / or mixtures of these.

These components in the aforementioned proportions have a greater effectiveness and prolonged effect in the fight of ticks, guaranteeing control in adults, larvae, nymphs and eggs, especially in adult females by altering their oviposition efficiency, reducing and eliminating the ability to deposit eggs. and also affecting the hatching of eggs and the survival of larvae.

Overview of! Chíicuague (Helíopsís Longipes)

Ei chíicuague (Helíopsís longipes S.F. Biake} is a perennial herb endemic to the Alvarez and Gorda mountains, a region where part of the territories of San Luis Potosí, Guanajuato and Guerétaro coincide, and is known as where its roots have traditional uses.

The genus Helíopsís belongs to the Heliantheae tribe of the Asteraceae family, with about 2,500 species grouped into 189 genera and divided into ten subtribes, including Zinnia, Phüactis, Sanvitalia, Acmella, Podac aenium, Squa opappus, Spilant es and Salmea, which make up the subtribe Zinníínae. Its members share in their morphology the presence of solitary chapters, conical or hemispherical receptacles, pistillate, fertile and persistent liguia flowers, fertile disc flowers and laterally compressed or angled achenes of the disc. The biggest difference that separates Helíopsís species from those of other genera is the combination of characters: the presence of fertile flowers in the disc, persistent and fertile leaved flowers, sessile and persistent corollas, thick achenes and absent vilane. Foliar margin is an important feature. Most species have jagged or semidentadas leaves, although some have them almost entirely or irregularly created. The texture of the leaf is generally firm, with the exception of annual plants that have usually thin and fragile leaves, the petiole has been used as a valuable character in the separation of H. helianthoides subspecies, both for its length and for its shape. Aleamides in the genus Hsiiapsis

Aicamides are synthesized by two different metabolic pathways. The acidic part originates from urs fatty acid of chain length which can be from eight to eighteen carbons, generally aliphatic or linear; This, when condensed with an amino acid and in concomitant decarboxylation, results in the production of an acamide. The amides, whose acidic chain is aiifatic, depending on the type of unsaturated bonds that they present can be separated into two groups: olefinic aicamides, with at least one double bond; and acetünic aicamides, with at least one tripie ligation. In the genus Heüopsis, only aiphytic and acetünic aicamides have been found.

An important inhibitory action of the aicamides of! chilcuague on the development of in vitro cultures of Escheríchia coli, Pseudomonas soíanacearu, Bacillus subtilis and Sacharomyces cerevisiae; as well as some phytopathogenic fungi such as Sclerotium cepivorum, Sclerotium rofísii and Fusarium oxysporu.

Afinin is the substance responsible for the biological effects of ehilcuague among which are considered activities of local anesthetic, flavoring, insecticide, fungicide and bactericide. It accumulates in its roots in large quantities (up to 1% of the total root weight).

Overview of Propole © [Propolis)

The propolis is a product of the hive formed by resins that bees collect from the buds of trees, which are mixed with the saliva, enzymes and other secretions of the bees. It is used as a natural protector against pathogenic microorganisms and fungi; protects against the cold during the winter and acts as a sealing material on the external and internal walls of the hive to reduce the entry of insects. This product is highly appreciated for its biological activities: bactericidal, antiviral, fungicidal, insecticidal anti-cancer, antioxidant, healing, immunostimulatory, anesthetic and analgesic among others. These features are related to its chemical composition, botanical origin, harvest time and the species of bee to collect.

Chemical composition or Propoüs

The chemical composition of the propo! Eo is very variable and complex, depending on the local flora near the site of the coimena. Its main components are resins and balms (50% to 55%), waxes (25% to 35%), voiaíiies oils (5% to 10%), pollen (2% to 6%), minerals and organic substances (5% to 8%), Among the latter, organic acids, phenolic acids, aromatic aldehydes, coumarins, phenolic compounds such as flavonoids (flavones, flavonoids, fiavanones, flavonoies) and minerals (Ai, Ag, Ba, B, Cr, Co, have been detected Cu, Sn, Fe; iVfg, Mn, Ni, Pb, Se, Si, Sr, Ti, V and Zn). It also contains vitamins A, B1, B2, B3 and B8. The group of compounds with biological activity most commonly evaluated in propolis extracts are known as phenolic compounds. Most of these occur in the form of flavonoids. Thus, the presence of the eight routine flavonoids, myricetin, quercetin, kaempfero !, apigenin, pinocembrina, chrysin and galangina, can be used as a marker to differentiate propolis from other products in the hive. The content of these eight flavonoids has been widely used as a parameter of the quality of propolis.

Other compounds identified in propolis are: dihydrofiavonones, derivatives of! benzylic aicohoi, benzaldehyde and benzoic acid, derivatives of cinnamic aicohoi, coumarins, phenolic triglycerides, other aromatic elements, a monoterpene, hexaterpenes, ítererpenes, sterols, a large amount of fatty acids, carbohydrates, poiisacársdos, vitamins and other compounds. Phenolic compounds constitute more than! 50% of its total weight

It has been shown that the effect of propolis on a wide variety of microorganisms and on some insects is dependent on chemical composition. Preparation of the extract of chiicuague {Heüopsis ¡angipes)

In the ursa modality of the present invention, the process for obtaining an extract of chiicuague is carried out as follows: a) Collection and selection of roots of H. íongipes.

b) Drying and grinding in a mill until obtaining fine poivo,

c) Extraction with water or with any other organic solvent in a ratio of 2 to 10 L / kg of chiicuague, with constant stirring for 18 to 30 hours at a temperature of 35 to 45 ° C. d) Filtration of the extract by means of a filter press or any other filtration method to remove suspended particles.

e) Evaporation or distillation of the extract obtained at a temperature of 45 to 75 ° C, until concentration,

f) Storage at 4 ° C until use or addition of the extract to the desired mixture.

Propolis extract preparation {PropoHs}

In one embodiment of the present invention, the process for obtaining an extract of propolis is: a) Selection and collection of resins directly from hives.

b) Cleaning of the selected resins.

c) Extraction with ethanol or with any other organic solvent in a proportion of 15% to 30% (by weight / volume), preferably using 70% to 95% ethanol.

d) acceleration of the extract for 5 to 10 days, with frequent agitation.

e) Filtration of the extract by means of a press filter or any other filtration method to remove suspended particles.

f) Storage in amber bottles, protected from light and at room temperature. Advantages

1. - E! Combined use of natural extracts for cons! of ticks, it guarantees the absence of resistance and a very low probability that they develop, since the combination of active metabolites in both extracts provides a mixture to which they will hardly adapt. It is expected that with this type of products, the development of resistances, (if it happens} takes prolonged periods of time,

2. - The composition showed a high percentage of control in terms of inhibition of oviposition and inhibition of hatching, as well as larval survival; It guarantees incremental results from generation to generation and could lead to the eradication of tick populations. 3.- In the composition of the propotropic extract (Propoiis) it causes an increase in permeation and a reduction in the potential of the membranes and cell walls, which contributes to reducing resistance to other toxic agents or constituents of the same extract or of other extracts, acting in synergy with the extract of chíícuague. In cases of extreme infestations, this property of the composition could be used to carry out treatment combined with chemical compounds to which the populations of ticks to combat are not yet fully resistant.

4.- The composition of the present invention is capable of controlling all the ticks of veterinary importance of the family Ixodidae (Ambíyomma spp., BoophHus spp., Dermacenior spp., Haemaphysaüs spp., HyaÍom a spp., Ixodes and Rhipicephalus} and of the Argasidae family (Ornithodorus spp. and Otobius spp.). Examples

EXAMPLE 1

Immersion tests on full females, performed with the composition and compared with water, showed control percentages of 70 to 93%. The test consists of immersing the full female for 1 minute in the composition and subsequently allowing it to perform oviposition in an environment of controlled temperature and humidity. 10 females are placed for each repetition. Subsequently, eggs deposited are weighed and their weight is compared against eggs deposited by females submerged in water, to calculate the% inhibition of oviposition. Once the females ovipositioned, the eggs are incubated until hatching and by counting under the microscope of the empty "shells", the percentage of inhibition of the hatching of Sos eggs from females treated with ia is calculated by comparison with the water control composition. Finally, the larvae that emerge from the eggs are incubated under controlled conditions and their survival rate is evaluated. With these three parameters the percentage of control is determined, which integrates the ability of the composition to inhibit oviposition, inhibit hatching and prevent larval survival, in a single value known as percent control; which the higher, indicates better results. Figure 1 shows the effect on oviposition.

Axis p! O 2

Trials with larvae using the Shaw test showed a range in mortality rates of 70.89 to 100%. Figure 2 shows the graph with results of 7 independent tests, using different concentrations of the composition within the range specified in the present invention; which consist of impregnating with 10m! of the composition a filter paper, place 300 larvae on the paper and cover them with another filter paper also impregnated with the composition. Controls are performed by impregnating the filter paper with water. The larvae remain in contact with the papers for 10 minutes and subsequently the papers are removed and the larvae are placed under conditions 5 temperature and humidity controlled to observe its mortality rate within 72 hours after the application of the composition. The larvae subjected to water treatment had 0% mortality.

Example 3 0 "Live m" trials with horses showed a decrease in infestation in a period of 72 hours after application of the formulation. The full females detached in their entirety and their mortality was 70%. Figures 3 and 4 show the effect on the wave and take respectively an equine specimen. The smposition was applied by spraying directly on the affected parts. S

Example 4

"In vivo" trials in cattle showed a decrease and infestation within 72 hours after application of the formulation, both in cases of high and low infestation. The full females detached in their entirety and their mortality was from! 90% Figures 0 and 5 show the effect of a bovine with a low infestation and a bovine with an alias infestation, respectively. The composition was applied by bath with water pump.

5 Description of the figures

Figure 1 shows two repetitions of! test with full females immersed in water (FiguralA) whose oviposition was considered as 100%. In Figure 1 B and 1C four repetitions of the test are shown with full females immersed in the composition at two different concentrations of the extracts, within the range specified in the claims of the present invention.

Figure 2 shows e! result of the survival of larvae treated with 7 different concentrations of the composition (within the range specified in the present invention), following the method of Sha. The result was a mortality range between 70.89 and 100%.

Figure 3 shows a series of photographs of the ear of a horse infested with ticks. Figure 3A shows the infested ear, Figure 3B shows the decrease in the presence of ticks after 24 hours of application of the composition and Figure 3C shows the release of 90% of the females at 72 hours after the application of the composition.

Figure 4 shows a series of photographs of the neck of a horse infested with ticks. A high population density of ticks attached to the horse's skin can be seen in Figures 4 A and B and in Figures 4C and 4D the detachment of 85% of the ticks can be seen 72 hours after the composition of the neck is applied. horse.

Figure 5 shows the legs of a bovine with low tick infestation, where Figure 5A shows the infested leg, Figure 5B shows the effects of detachment and mortality of females (due to the color change of the ticks, which are dead although they have not come off) 24 hours after applying the composition and Figure 5C shows the width 72 hours after applying the composition, showing 90% detachment and mortality of the ticks. 33 Figure 6 shows the legs of a bovine with a tick infestation, where the figure

6A shows the infested leg, figure 68 shows the effects of detachment and mortality of females (evident by the color change of the ticks, which are dead even if they have not detached} 24 hours after applying the composition and figure 6C shows he is 72 hours after the composition is applied, evidencing a 80% mortality and detachment of the ticks.

Claims

1. A composition based on natural extracts for the control and / or elimination of ticks, characterized in that it comprises:

a) A natural extract of H & iiopsis longipes;

b) A natural extract of Propolis and

c) A dermal vehicle

2. A composition according to claim 1, characterized in that it comprises:

a) A natural extract of Heliopsis longipes from 50 to 65% v / v

b) A natural extract of Propolis from 5 to 20% v / v and

c) A 30% dermal vehicle v / v

All this must add up to 100%.

3. A composition according to claim 1, characterized in that the dermal vehicle is selected from glycerin, mineral oil or petrolatum and / or mixtures thereof.

4. A composition according to claim 1, characterized in that it is used for the control of populations of adults, nymphs, pupae and tick eggs.

5. A composition according to claim 1, characterized in that it is used for the control and possible eradication of ticks, especially in adult females altering their ability to oviposition, decreasing also affecting the hatching of the eggs and the survival of the larvae.

6. A composition according to claim 1, characterized in that it is used for e! control and / or elimination of all ticks of veterinary importance of the family Ixodidae (Amblyomma spp., Boophilus spp., D rmacenior spp., Hae aphysaiis spp., Hyalomrna spp., xodes and Rhipic & phaius) and of the family Argasidae ( Or sthodorus spp. And Qtobius spp.).

7. A composition according to claim 1, characterized in that it is used for the control and / or elimination of ticks, especially AmbSyomma cajennensi, Ambfyo ma americanum, Amblyomma macuiatum, AmbSyomma ebraeu, Ambiyomma varíegatu, Boophíius micropíus, Boophiius decoíoraíu, Boophiiat annioiusus, Boophiius decoioiusus, Boophiius decoioraius , Dermacentor margínalas, Dermacentor pictus, Dermacentor reticuiatus, Dermacentor nitens, Dermacentor variabais, ixodes scapuiarís and Rhipicep aius blood.

8. A composition according to claim 1, characterized in that it can be applied to cattle, sheep, goats, horses and domestic animals.

S. A composition according to claim 1, characterized in that the chiicuague extract (Heiiopsss íongipes) has an effect of paralysis and toxicity on ticks, mainly attacking adult populations.

10. A composition according to claim 1, characterized in that e! Propolis extract (Propoiis) causes an increase in permeability and a reduction in the potential of cell membranes and walls, contributing to decrease resistance to other toxic agents or constituents of the same extract or other extracts, acting in synergy with Chiicuague extract.

11. A composition according to claim 1, characterized in that the active substance responsible for the biological effects of chiicuague is afinine alkamide.

12. A composition according to claim 1, characterized in that the active substances responsible for the insecticidal effects of propolis are mainly coumarins.

13. A process for obtaining a chiunguague extract characterized in that it comprises the following steps:

a) Collection and selection of roots of H. Íongipes.

b) Drying and grinding in a mill until fine dust is obtained.

c) Extraction with water or with any other organic solvent in a ratio of 2 to 10

L / kg of chiicuague, with constant agitation for 18 to 30 hours at a temperature of 35 to 45 ^e C. d) Filtration of the extract by means of a filter press or any other filtration method to remove suspended particles,

e) Evaporation or distillation of! extract obtained at a temperature of 45 to 65 ⁰ C until concentration.

14. A process for obtaining a propolis extract characterized in that it comprises the following steps:

a) Selection and collection of resins directly from bee hives. b) Cleaning of resins to remove wax and other organic particles

c) Mix of the selected resins.

d) Extraction with ethanol or with any other organic solvent in a proportion of 15% to 30% (by weight / volume), preferably using 70% to 95% etanoi.

e) Maceration of the extract for 5 to 10 days, with frequent agitation.

f) Filtration of the extract by means of a filter press or any other filtration method to remove suspended particles.

15. The process according to claims 13 and 14, characterized in that the composition was formed with: a} 50% to 85% natural extract of Heliopsis longipes; b) 5% to 20% of natural extract of Propoiis and c) 30% of a dermal vehicle, all this must add up to 100%.